Fig. 4.

Construction of pyroptosis model using primary hepatocytes of grass carp. Cell viability (A), Hoechst/PI staining (B), and quantification (C) after 100 ng/mL LPS treatment for 4 h, followed by different concentrations of Nig treatment for 0.5 h. Gene expression of NLRP3, ASC, caspase-1, GSDME, and IL-1β (D); NLRP3 immunofluorescence staining (E) and quantification (F). Protein expression of cleaved-caspase-1, GSDME, N-GSDME, IL-1β (G) and quantification (H) in these two groups. N: Nucleus. The results were expressed as mean ± SD of 3 or 6 independent observations (WB: n = 3). ^a–fValues having different letters are significantly different (P < 0.05); ^**P < 0.01, ^***P < 0.001. Nig, nigericin sodium salt; Con, control; NLRP3, NOD-like receptor thermal protein domain associated protein 3; ASC, apoptosis-associated speck-like protein containing a CARD; CASP-1, cysteinyl aspartate specific proteinase-1; GSDME, gasdermin E; IL-1β, interleukin-1β